RNA-seq and de novo assembly protocol

The RNA-seq and de novo assembly was performed as previously described [21 (link),22 (link),23 (link)]. Briefly, total RNA was extracted from the sampled leaf tissues using the EASYspin Plus Complex Plant RNA Kit (Aidlab Biotech, Beijing, China). Ribosomal RNAs were removed by the TruSeq RNA Sample Prep Kit (Illumina, San Diego, CA, USA). An RNA library was constructed using the TruSeq RNA Sample Prep Kit according to the manufacturer’s protocol (Illumina, San Diego, CA, USA), followed by sequencing on the Illumina HiSeq X-ten platform (Biomarker Technologies, Beijing, China). Low-quality reads were filtered and adapters of the paired-end raw reads were trimmed using the CLC Genomics Workbench 9.5 software (QIAGEN, Hilden, Germany). The clean reads were de novo assembled into contigs using the Trinity v2.3.2 program (Broad Institute and the Hebrew University of Jerusalem, Cambridge, Massachusetts and Jerusalem, USA and Israel) [22 (link)].